AFFINITY ISOLATION & I-DIRT MS ANALYSIS OF E COLI O157:H7 SAKAI RNA POL COMPLEX

亲和隔离

• 批准号: 8169142
• 负责人: Brian T Chait
• 金额: $ 0.12万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8169142
• 关键词: Affinity; Bacteria; Code; Complex; Computer Retrieval of Information on Scientific Projects Database; DNA-Directed RNA Polymerase; Escherichia coli; Escherichia coli O157; Funding; Genes; Genome; Grant; Institution; Laboratories; Manuscripts; Mass Spectrum Analysis; Multienzyme Complexes; Proteins; Publishing; RNA; Research; Research Personnel; Resources; Source; Techniques; Transcription Initiation; Transcriptional Regulation; United States National Institutes of Health; Work; pathogenic Escherichia coli

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Bacteria contain a single multi-subunit RNA polymerase that is responsible for the synthesis of all RNA. Previous studies on the Escherichia coli K-12 laboratory strain had identified a group of effector proteins that interact directly with RNA polymerase to modulate the efficiency of transcription initiation, elongation or termination. Here we have used a rapid affinity isolation technique to isolate RNA polymerase from the pathogenic Escherichia coli O157:H7 Sakai strain. We analysed the RNA polymerase enzyme complex using mass spectrometry and identified associated proteins. Although Escherichia coli O157:H7 Sakai contains more than 1,600 genes not present in the K-12 strain, many of which are predicted to be involved in transcription regulation, all of the identified proteins in this study were coded on the 'core' Escherichia coli genome. A Manuscript describing this work was published: Affinity isolation and I-DIRT mass spectrometric analysis of the Escherichia coli O157:H7 Sakai RNA polymerase complex. Lee DJ, Busby SJ, Westblade LF, Chait BT. J Bacteriol. 2008 Feb;190(4):1284-9.

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 细菌含有一个负责合成所有RNA的多亚单位RNA聚合酶。以前对大肠杆菌K-12实验室菌株的研究已经确定了一组直接与RNA聚合酶相互作用的效应蛋白，以调节转录的启动、延伸或终止的效率。在这里，我们使用了一种快速亲和分离技术来分离致病性大肠杆菌O157：H7酒井株的RNA聚合酶。我们用质谱仪分析了RNA聚合酶复合体，并鉴定了相关蛋白。虽然大肠杆菌O157：H7 Sakai含有1,600多个K-12菌株没有的基因，其中许多被预测参与转录调控，但本研究中所有已鉴定的蛋白质都编码在大肠杆菌的核心基因组上。一份描述这项工作的手稿已经出版： 大肠杆菌O157：H7 Sakai RNA聚合酶复合体的亲和分离和I-污物质谱分析。Lee DJ，Busby SJ，WestBlade LF，Chait BT。细菌素J。2008年2月；190(4)：1284-9。